Compositions and methods for inhibiting kinases

ABSTRACT

The present invention provides compounds for the prevention or treatment of cancer or a bacterial or viral infection. Additionally, the present invention provides compositions and methods for using these compounds and compositions in the prevention or treatment of cancer or a bacterial or viral infection in a subject.

PRIORITY CLAIM

This application is a continuation of U.S. patent application Ser. No.15/805,693, now issued as U.S. Pat. No. 10,118,923, filed Nov. 7, 2017,which is a continuation of U.S. patent application Ser. No. 15/136,497,now issued as U.S. Pat. No. 9,828,370, filed Apr. 22, 2016, which claimsthe benefit of, and priority to, U.S. Provisional Patent Application No.62/182,955, filed Jun. 22, 2015, and U.S. Provisional Patent ApplicationNo. 62/151,659, filed Apr. 23, 2015. The disclosure of each of theseapplications is hereby incorporated by reference herein in its entirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under Contract Nos.NS103695 and AI103982, awarded by The National Institutes of Health. Thegovernment has certain rights in the invention.

BACKGROUND OF THE INVENTION

Anti-Infective Agents

Abelson-family tyrosine kinases (ATKs) are host targets that can beinhibited by small molecules (ATKis) to create anti-cancer andanti-infective therapeutics. When applied to treat infectious diseasesin vitro and in vivo, more than 20 human bacterial viral, fungal andparasitic pathogens have been shown to be susceptible to ATK inhibition.Host ATKs are co-opted by a pathogen to enter, reproduce or exit hostcells, accounting for the antimicrobial effects of ATKis. Because thetargets of these therapeutics to treat infectious disease overlap withthe targets for certain types of cancer, the same medications forinfectious disease can be applied to treat cancer at the same dose.

For infectious disease, this strategy has been applied to treat thecause of Progressive Multifocal Leukoencephalopathy (PML) a fatal braininfection that arises in chronically immunosuppressed populationsincluding people with HIV1 infection, patients on chronicimmunosuppressive therapy such as corticosteriods for organ transplant,patients with cancer and/or autoimmune diseases (such as rheumatoidarthritis, psoriasis, and lupus erythematosis), and patients ontherapies that depress the immune response (e.g., efalizumab,belatacept, rituximab, natalizumab, infliximab, among others). PML is afatal condition caused by the lytic infection of JC polyomavirus (JCV)in the brain. However, the brain-infective form of the virus is notacquired by transmission from an infected patient. Instead,brain-infective JCV is formed by genomic rearrangement of thenon-pathogenic form of the virus that resides outside of the brain in apersistently infective state within a patient. This non-pathogenic, orarchetype JCV, which is detected in the kidney, genitourinary tract andbone marrow, is kept in check by the host cellular immune response. Thetransformation to the brain-infective form occurs when a patientundergoes a sustained disruption of cellular immunity, which could becaused by either immune-suppressing drugs to treat autoimmune disease(e.g. MS) or by development of clinical AIDS following HIV infection. Inthe context of immunosuppression, JCV rearranges its non-coding controlregion (NCCR), followed by mutations in the viral capsid protein VP1.Rearranged NCCR is thought to enable the virus to replicate in a widerrange of cells, while the subsequent mutations in VP1 enable viral entrythrough a broader range of receptors found on the surface of cellsoutside the genitourinary tract. Neither the pathway to the brain northe carrier enabling JCV to enter the brain is definitively known, butit is a slow process that takes years to complete. A characteristicmarker of PML is the appearance of viral DNA in the cerebrospinal fluid(CSF). In a typical clinical case, a patient is negative for JCV DNAwithin 2 months of a diagnosis, after which JCV DNA is readily found inthe CSF. Thus, the process of central nervous system (CNS) entry is veryslow, but once virus enters the CNS, progression to disease isrelatively rapid.

Lytic infection of JCV in the brain causes irreversible damage to neuraltissue. Thus, it would be desirable to clear JCV from a patient early inthe course of treatment. An ATKi could be used alongside therapies likeTysabri for MS, to clear JCV at the initiation of an immune-suppressingtreatment.

Proof-of-concept trials related to a PML antiviral program to treat JCVinfection with the marketed drug Gleevec, a well-known anticancerAbl-kinase inhibitor, were conducted. Gleevec was very useful fordefining the mechanism of action, but it rapidly became clear that thehuman steady-state (SS) concentration of Gleevec cannot sustain anefficacious dose for an antiviral purpose (Table 1). The steady-statetrough concentration, C_(min) ^(SS), is just 2.3-fold higher than theEC₅₀ of Gleevec against JCV in cell culture (Table 1). Typically, thisratio should be 4- to 9-fold above the EC₅₀ value for a safe andeffective antiviral agent.

Improved agents for treating JCV and other infectious agents are needed.

Anti-Cancer Agents

Chronic Myeloid Leukemia (CML) is a myeloproliferative neoplasm with anincidence of 1-2 cases per 100,000 persons, and accounts for ≈15% ofnewly diagnosed cases of leukemia in adults. Pathogenesis of CML islinked to the fusion of the Abelson murine leukemia (ABL) gene onchromosome 9 with the breakpoint cluster region (BCR) gene on chromosome22, resulting in expression of fusion protein, termed BCR-ABL. BCR-ABLis a constitutively active tyrosine kinase that promotes growth andreplication through downstream pathways such as RAS, RAF, JUN kinase,MYC, and STAT. The consequences of BCR-ABL expression create acytokine-independent cell cycle with aberrant apoptotic signals inresponse to cytokine withdrawal. The development of small molecule ATKisthat potently interfere with the interaction between BCR-ABL and ATPwere shown to block cellular proliferation of the malignant clone. This“targeted” approach was found to dramatically alter the natural historyof the disease, improving 10-year overall survival (OS) from ≈20% to>80%.

Three such targeted therapies have been approved for first linetreatment of CML: imatinib (Gleevec®), nilotinib (Tasigna®) anddasatinib (Sprycel®). Gleevec, the first of these agents to reachmarket, displayed a remarkable 81% event-free survival rate and a 93%overall survival rate when CML-only related deaths were considered. But,8-year follow-up studies revealed that only 55% of patients remained ontherapy, indicating that additional options were needed to handletreatment failure and improve tolerability of Gleevec as a chronicmedication. Treatment failures were often linked to the development ofGleevec resistance arising from secondary mutations in the Abl-kinasedomain of the fusion protein, resulting in a loss of Gleevec potency andrelapse of disease. Sprycel, a dual Abl- and SRC-kinase inhibitor, wasthe second approved agent and a much more potent inhibitor of BCR-ABL.Sprycel induces more rapid responses and suppression of fusion proteinexpression at much earlier timepoints than Gleevec. But, as a SRCinhibitor, significant side effects counterbalanced the value of theenhanced response profile that Sprycel displayed. Tasigna was developedto directly address Gleevec resistance, and, early in its development,demonstrated potent inhibitory activity against many clinically relevantBCR-ABL mutants that no longer responded to Gleevec therapy (Table 3).Like Sprycel, Tasigna was as much as 20× more potent as an inhibitor ofBCR-ABL relative to Gleevec (Table 3). However, Tasigna's potency wasaccompanied by a side effect profile with severe adverse event frequencysimilar to Sprycel, limiting its utility as a chronic medication.

Thus, despite the success of targeted therapies against BCR-ABL,comorbidities and toxicities are a dominant issue in the use of thesefrontline ATKis for some patients. Patients at risk of developingpleural effusions, for example, such as patients with a history of lungdisease (e.g., COPD), cardiac disease (e.g., congestive heart failure orpulmonary arterial hypertension) or patients with uncontrolledhypertension, make Sprycel a poor choice. Sprycel also inhibits plateletfunction, so patients taking anticoagulants could be at risk forbleeding complications while on Sprycel. Tasigna is associated withhyperglycemia and therefore could be detrimental if used in patientswith uncontrolled diabetes. Tasigna may also prolong the QT interval andtherefore may be contraindicated in patients with cardiac complications,although longer-term follow-up studies appear to be needed to confirmthis observation. The non-linear accumulation of Tasigna that occurswhen taken in the context of a fatty diet also places patients at riskfor reaching severe dose-limiting toxicities, requiring fasting beforeand after dosing with Tasigna for 2 hours; as Tasigna is given 2×/day,this means patients on Tasigna are fasting as much as 8 hours out ofevery 24. By contrast, while Gleevec is associated with some severe AEs(e.g., leukopenia and cytopenia), its most prominent side effect isperipheral edema, which can be medically managed. Ponatinib, the mostrecent ATKi to reach market and which addresses nearly all Gleevecresistance, including the T315I ‘gatekeeper’ mutation, was subsequentlyfound to cause a severe blood clotting syndrome and a narrowing of bloodvessels after 24 months of therapy. As a result, ponatinib's utility hasbeen severely restricted as a third-line treatment when no other optionsexist. Bosutinib, another recently approved Abl-SRC dual inhibitor, isconsidered second-line in the context of imatinib-resistance withpotency that is similar to Sprycel.

Similarly, GastroIntestinal Stromal Tumors (GIST), the most commonmesenchymal tumors originating in the digestive tract, have acharacteristic morphology and are generally positive for CD117 (c-kit)and are primarily caused by activating mutations in the KIT or PDGFRa,both protein kinases in the Abelson-kinase family which are susceptibleto treatment with ATKis. Just as found in the case of BCR-Abl associatedcancers, KIT and/or PDGFRa associated GIST treated with frontline TKIslike imatinib can eventually develop resistance to therapy through theformation of secondary mutations in either KIT or PDGFRa; imatinib alsodisplays poor response rates in KIT exon 9 and wildtype associatedtumors, both of which are more responsive to higher affinity ATKis likesunitinib and regorafenib. Given the lower overall response rates ofsunitinib and regorafenib relative to imatinib in front line therapy,the development of new agents with the broadest application in themanner of imatinib offers a likelihood of treatment success that willexceed the higher affinity agents like sunitinib and regorafenib.

SUMMARY OF THE INVENTION

In one aspect, the invention provides compounds represented by generalformula (I) or a pharmaceutically acceptable salt thereof:

wherein, independently for each occurrence,R¹ is selected from hydrogen or lower alkyl; andCy¹ is selected from substituted or unsubstituted aryl, substituted orunsubstituted heteroaryl, and substituted or unsubstituted heterocyclyl,provided that Cy¹ is not unsubstituted pyrid-4-yl.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A shows the effect of imatinib and various compounds of thepresent invention on T1 cells containing the GIST-related cKit 557-558deletion.

FIG. 1B shows the effect of imatinib and various compounds of thepresent invention on 430 cells containing the GIST-related cKit 557-558deletion.

FIG. 1C shows the effect of imatinib and various compounds of thepresent invention on T1 cells containing the GIST-related cKit 557-558deletion and the D816E mutation.

FIG. 1D shows the effect of imatinib and various compounds of thepresent invention on T1 cells containing the GIST-related cKit 557-558deletion and the D820Y mutation.

FIG. 1E shows the effect of imatinib and various compounds of thepresent invention on T1 cells containing the GIST-related cKit 557-558deletion and the A829P mutation.

FIG. 1F shows the effect of imatinib and various compounds of thepresent invention on ACC-430 cells containing the GIST-related cKit557-558 deletion and the V654A and N680K mutations.

FIG. 1G shows the effect of imatinib and various compounds of thepresent invention on 430 cells containing the GIST-related cKit 557-558deletion.

FIG. 2 depicts a Western blot showing the effect of imatinib and variouscompounds of the present invention on c-Kit phosphorylation, AKTphosphorylation, and MAPK phosphorylation.

FIG. 3 shows the suppression of tumor growth for K562 cell BCR-Ablxenografts in NSG mice as a model of Chronic Myelogenous Leukemia.

DETAILED DESCRIPTION OF THE INVENTION

In one aspect, the invention provides compounds represented by formula(I) or a pharmaceutically acceptable salt thereof:

wherein, independently for each occurrence,R¹ is selected from hydrogen or lower alkyl; andCy¹ is selected from substituted or unsubstituted aryl, substituted orunsubstituted heteroaryl, and substituted or unsubstituted heterocyclyl,provided that Cy¹ is not unsubstituted pyrid-4-yl.

In certain embodiments, Cy¹ is selected from:

wherein, independently for each occurrence,R² and R³ are selected from hydrogen, alkyl, amino, monoalkylamino,dialkylamino, cycloalkyl, halo, cyano, alkoxy, —C(O)OH, and—C(O)N(R⁴)(R⁴);n is 1, 2, 3 or 4;X is C(R⁴)₂, S, O, or NR⁴;R⁴ is selected from hydrogen and substituted or unsubstituted alkyl,aralkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, heteroaralkyl,cycloalkylalkyl, or heterocyclylalkyl.

In certain embodiments, Cy¹ is selected from:

wherein, independently for each occurrence,R¹ is selected from hydrogen or lower alkyl;R² and R³ are selected from hydrogen, alkyl, amino, monoalkylamino,dialkylamino, cycloalkyl, halo, cyano, alkoxy, —C(O)OH, and—C(O)N(R⁴)(R⁴);R⁴ is selected from hydrogen and substituted or unsubstituted alkyl,aralkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, heteroaralkyl,cycloalkylalkyl, or heterocyclylalkyl.

In certain other embodiments, Cy¹ is not substituted or unsubstitutedpyrid-4-yl.

In certain embodiments, Cy¹ is 5-membered heteroaryl, aryl orheterocyclyl.

In certain embodiments, Cy¹ is selected from:

In certain embodiments, Cy¹ is selected from:

In certain embodiments, Cy¹ is selected from:

In certain embodiments, Cy¹ is selected from:

In certain preferred embodiments, R¹ is methyl, e.g., —CH₃, —CDH₂,—CD₂H, or —CD₃.

In one aspect, the invention provides a pharmaceutical compositioncomprising a compound as disclosed herein.

In certain embodiments, the pharmaceutical composition further comprisesone or more pharmaceutically acceptable excipients.

In one aspect, the invention provides a compound or composition, asdisclosed herein, for conjoint administration with one or more compoundsindependently selected from central nervous system drugs, such asCNS/respiratory stimulants, analgesics, narcotic agonists, narcoticantagonists, nonsteroidal anti-inflammatory/analgesic agents,behavior-modifying agents, tranquilizers/sedatives, anesthetic agents,inhalants, narcotics, reversal agents, anticonvulsants, skeletal musclerelaxants, smooth muscle relaxants, cardiovascular agents, inotropicagents, antiarrhythmic drugs, anticholinergics, vasodilating agents,agents used in treatment of shock, alpha-adrenergic blocking agents,beta-adrenergic blocking agents, respiratory drugs, bronchodilators,sympathomimetics, antihistamines, antitussives, agents for urinaryincontinence/retention, urinary alkalinizers, urinary acidifiers,cholinergic stimulants, agents for urolithiasis, gastrointestinalagents, antiemetic agents, antacids, histamine H2 antagonists,gastromucosal protectants, proton pump inhibitors, appetite stimulants,GI antispasmodics-anticholinergics, GI stimulants, laxatives, saline,lubricant, surfactant, antidiarrheals, hormones/endocrine/reproductiveagents, sex hormones, anabolic steroids, posterior pituitary hormones,adrenal cortical steroids, glucocorticoids, antidiabetic agents, thyroiddrugs, thyroid hormones, endocrine/reproductive drugs, prostaglandins,antiinfective drugs, antiparasitics, anticoccidial agents, antibiotics,anti-tuberculosis, aminocyclitols, cephalosporins, macrolides,penicillins, tetracyclines, lincosamides, quinolones, sulfonamides,antibacterials, antifungal agents, antiviral agents, blood modifyingagents, clotting agents, anticoagulants, erythropoietic agents,antineoplastics/immunosuppressives, alkylating agents, antidotes,bone/joint agents, dermatologic agents (systemic), vitamins andminerals/nutrients, systemic acidifiers, systemic alkalinizers,anti-cancer agents, and anti-viral agents.

In another aspect, the invention provides a use of a compound to treatProgressive Multifocal Leukoencephalopathy (PML), e.g., in multiplesclerosis (MS) patients on Tysabri, immunosuppressed patients (includingpatients with HIV1 infection), patients on chronic immunosuppressivetherapy such as corticosteroids for organ transplant, patients withcancer and/or an autoimmune disease (such as rheumatoid arthritis,psoriasis, and lupus erythematosis), and patients on therapies thatdepress the immune response (e.g., efalizumab, belatacept, rituximab,natalizumab, infliximab, among others).

In another aspect, the invention provides a use of a compound orcomposition as disclosed herein for altering activity of one or moreATKs in a mammal, such as a human.

In another aspect, the invention provides a use of a compound orcomposition as disclosed herein for altering the function of c-Abl1and/or c-Abl2 or any protein comprising a kinase domain of c-Abl1 and/orc-Abl2 in a mammal, such as a human.

In another aspect, the invention provides a use of a compound orcomposition as disclosed herein for altering the function of c-Kit orany protein comprising a kinase domain of c-Kit in a mammal, such as ahuman.

In another aspect, the invention provides use of a compound orcomposition as disclosed herein for inhibiting PGDFRa or PDGFRb or anyprotein comprising a kinase domain of PDGRFa or PDGRFb.

In another aspect, the invention provides use of a compound orcomposition as disclosed herein for treatment of cardiovascularabnormalities such as pulmonary arterial hypertension (PAH), HIV-relatedKaposi's Sarcoma, Idiopathic Pulmonary Fibrosis (IPF), Diffuse CutaneousSystemic Sclerosis or Rheumatoid Arthritis.

In another aspect, the invention provides use of a compound orcomposition as disclosed herein for inhibiting mutated c-Kit having oneor more mutations associated with Gastrointestinal Stromal Tumor (GIST)such as those in Exon 11 (557-558 deletion), Exon 11 557-558 deletion incombination with a D186E mutation, Exon 11 557-558 deletion incombination with a D820Y mutation, Exon 11 557-558 deletion incombination with an A829P mutation, Exon 11 557-558 deletion incombination with V654K mutation, and Exon 11 557-558 deletion incombination with V654K and N680K. Mutation(s) in Exon 9 may also beaccessible to ATKi treatment.

In another aspect, the invention provides a method of treating a mammalsuffering from cancer, comprising administering to the mammal aneffective amount of a compound or composition as disclosed herein.

In another aspect, the invention provides a method of treatingGastrointestinal Stromal Tumors (GIST), the most common mesenchymaltumors originating in the digestive tract, which have a characteristicmorphology, are generally positive for CD117 (c-kit) and are primarilycaused by activating mutations in the KIT or PDGFRa, both proteinkinases in the Abelson-kinase family which are susceptible to treatmentwith ATKis, comprising administering a compound or composition, asdisclosed herein, to the subject.

In another aspect, the invention provides a method for preventing ortreating a bacterial infection or a viral infection in a subject,comprising administering a compound or composition, as disclosed herein,to the subject.

In certain embodiments, the invention provides for a method ofpreventing or treating a bacterial infection, e.g., a bacterialinfection caused by Pseudomonas aeruginosa, Chlamydia trachomatis,Escherichia coli, Helicobacter pylori, Listeria monocytogenes,Salmonella typhimurium, Shigella flexneri, or Mycobacteriumtuberculosis.

In certain embodiments, the invention provides for a method ofpreventing or treating a viral infection, e.g., a viral infection causedby a Vaccinia virus, a variola virus, a polyoma virus, a Pox virus, aHerpes virus, a cytomegalovirus (CMV), a human immunodeficiency virus,JC virus, JC polyomavirus (JCV), BK polyomavirus (BKV), Simian virus 40(SV40), Monkeypox virus, Ebola virus, Marburg virus, Bunyavirus,Arenavirus, Alphavirus (e.g., Venezuelan equine encephalitis (VEE) orWestern equine encephalitis (WEE)), Flavivirus, West Nile virus orCoronavirus (e.g., SARS).

In certain embodiments, the invention provides a method of preventing ortreating a viral infection, where the viral infection is a lyticinfection of JCV in the brain.

In certain embodiments, the invention provides a method of preventing ortreating a bacterial or viral infection, where the subject is a human.

In another aspect, the invention provides a method of preventing ortreating a bacterial or viral infection, where the compound orcomposition, as disclosed herein, is administered orally, nasally,buccally, sublingually, intravenously, transmucosally, rectally,topically, transdermally, subcutaneously, by inhalation, orintrathecally.

Compounds

Compounds of the invention include compounds of Formula I as disclosedabove and their salts (including pharmaceutically acceptable salts).Such compounds are suitable for the compositions and methods disclosedherein.

Definitions

The term “alkyl” refers to the radical of saturated aliphatic groups,including straight-chain alkyl groups, and branched-chain alkyl groups.In preferred embodiments, a straight chain or branched chain alkyl has30 or fewer carbon atoms in its backbone (e.g., C₁-C₃₀ for straightchains, C₃-C₃₀ for branched chains), and more preferably 20 or fewer. Incertain embodiments, alkyl groups are lower alkyl groups, e.g. methyl,ethyl, n-propyl, i-propyl, n-butyl and n-pentyl.

Moreover, the term “alkyl” (or “lower alkyl”) as used throughout thespecification, examples, and claims is intended to include both“unsubstituted alkyls” and “substituted alkyls”, the latter of whichrefers to alkyl moieties having substituents replacing a hydrogen on oneor more carbons of the hydrocarbon backbone. In certain embodiments, astraight chain or branched chain alkyl has 30 or fewer carbon atoms inits backbone (e.g., C₁-C₃₀ for straight chains, C₃-C₃₀ for branchedchains). In preferred embodiments, the chain has ten or fewer carbon(C₁-C₁₀) atoms in its backbone. In other embodiments, the chain has sixor fewer carbon (C₁-C₆) atoms in its backbone.

The term “alkenyl”, as used herein, refers to an aliphatic groupcontaining at least one double bond and is intended to include both“unsubstituted alkenyls” and “substituted alkenyls”, the latter of whichrefers to alkenyl moieties having substituents replacing a hydrogen onone or more carbons of the alkenyl group. Such substituents may occur onone or more carbons that are included or not included in one or moredouble bonds. Moreover, such substituents include all those contemplatedfor alkyl groups, as discussed below, except where stability isprohibitive. For example, substitution of alkenyl groups by one or morealkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups iscontemplated. In preferred embodiments, a straight chain or branchedchain alkenyl has 1-12 carbons in its backbone, preferably 1-8 carbonsin its backbone, and more preferably 1-6 carbons in its backbone.Examplary alkenyl groups include allyl, propenyl, butenyl,2-methyl-2-butenyl, and the like.

The term “alkynyl”, as used herein, refers to an aliphatic groupcontaining at least one triple bond and is intended to include both“unsubstituted alkynyls” and “substituted alkynyls”, the latter of whichrefers to alkynyl moieties having substituents replacing a hydrogen onone or more carbons of the alkynyl group. Such substituents may occur onone or more carbons that are included or not included in one or moretriple bonds. Moreover, such substituents include all those contemplatedfor alkyl groups, as discussed above, except where stability isprohibitive. For example, substitution of alkynyl groups by one or morealkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups iscontemplated. In preferred embodiments, an alkynyl has 1-12 carbons inits backbone, preferably 1-8 carbons in its backbone, and morepreferably 1-6 carbons in its backbone. Exemplary alkynyl groups includepropynyl, butynyl, 3-methylpent-1-ynyl, and the like.

The term “aralkyl”, as used herein, refers to an alkyl group substitutedwith one or more aryl groups.

The term “aryl”, as used herein, include substituted or unsubstitutedsingle-ring aromatic groups in which each atom of the ring is carbon.Preferably the ring is a 5- to 7-membered ring, more preferably a6-membered ring. Aryl groups include phenyl, phenol, aniline, naphthyl,biphenyl, anthracenyl and the like.

The term “cycloalkyl”, as used herein, refers to the radical of asaturated aliphatic ring. In preferred embodiments, cycloalkyls havefrom 3-10 carbon atoms in their ring structure, and more preferably from5-7 carbon atoms in the ring structure. Suitable cycloalkyls includecycloheptyl, cyclohexyl, cyclopentyl, cyclobutyl and cyclopropyl.

The terms “cycloalkyl” and “cycloalkenyl” refer to cyclic hydrocarbongroups of 3 to 12 carbon atoms.

The terms “halogen”, “halide” and “halo”, as used herein, mean halogenand include fluoro, chloro, bromo and iodo.

The terms “heterocyclyl”, “heterocycle”, “heterocyclo” and“heterocyclic” refer to substituted or unsubstituted non-aromatic ringstructures, preferably 3- to 10-membered rings, more preferably 3- to7-membered rings, whose ring structures include at least one heteroatom,preferably one to four heteroatoms, more preferably one or twoheteroatoms. The heterocyclic group may be attached at any heteroatom orcarbon atom of the ring or ring system. Exemplary monocyclicheterocyclic groups include pyrrolidinyl, pyrrolyl, pyrazolyl, oxetanyl,pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl,oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl,thiazolidinyl, isothiazolyl, isothiazolidinyl, furyl, tetrahydrofuryl,thienyl, oxadiazolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-oxoazepinyl, azepinyl,4-piperidonyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl,tetrahydropyranyl, morpholinyl, thiamorpholinyl, thiamorpholinylsulfoxide, thiamorpholinyl sulfone, 1,3-dioxolane andtetrahydro-1,1-dioxothienyl, triazolyl, triazinyl, and the like.Exemplary bicyclic heterocyclic groups include indolyl, benzothiazolyl,benzoxazolyl, benzodioxolyl, benzothienyl, quinuclidinyl, quinolinyl,tetra-hydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl,indolizinyl, benzofuryl, chromonyl, coumarinyl, benzopyranyl,cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, furopyridinyl (suchas furo [2,3-c] pyridinyl, furo [3,2-b] pyridinyl] or furo [2,3-b]pyridinyl), dihydroisoindolyl, dihydroquinazolinyl (such as3,4-dihydro-4-oxo-quinazolinyl), tetrahydroquinolinyl and the like.Exemplary tricyclic heterocyclic groups include carbazolyl, benzindolyl,phenanthrolinyl, acridinyl, phenanthridinyl, xanthenyl and the like.

The term “heteroalkyl”, as used herein, refers to a saturated orunsaturated chain of carbon atoms including at least one heteroatom(e.g., O, S, or NR⁴, such as where R⁴ is H or lower alkyl).

The term “heteroaryl” includes substituted or unsubstituted aromaticsingle ring structures, preferably 5- to 7-membered rings, morepreferably 5- to 6-membered rings, whose ring structures include atleast one heteroatom (e.g., O, N, or S), preferably one to four or oneto 3 heteroatoms, more preferably one or two heteroatoms. When two ormore heteroatoms are present in a heteroaryl ring, they may be the sameor different. The term “heteroaryl” also includes polycyclic ringsystems having two or more cyclic rings in which two or more carbons arecommon to two adjoining rings wherein at least one of the rings isheteroaromatic, e.g., the other cyclic rings can be cycloalkyls,cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls.Preferred polycyclic ring systems have two cyclic rings in which both ofthe rings are aromatic. Exemplary heteroaryl groups include pyrrolyl,pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl,isothiazolyl, furyl, thienyl, oxadiazolyl, pyridinyl, pyrazinyl,pyrimidinyl, quinolinyl, pyridazinyl, triazolyl, triazinyl, and thelike.

The term “alkoxy” is intended to mean an alkyl radical, as definedherein, attached directly to an oxygen atom. Some embodiments are 1 to 5carbons, some embodiments are 1 to 4 carbons, some embodiments are 1 to3 carbons and some embodiments are 1 or 2 carbons. Examples includemethoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy,5-isobutoxy, sec-butoxy, and the like.

The term “heteroatom”, as used herein, means an atom of any elementother than carbon or hydrogen. Preferred heteroatoms are nitrogen,oxygen, and sulfur.

The term “substituted” refers to moieties having substituents replacinga hydrogen on one or more carbons of the backbone. It will be understoodthat “substitution” or “substituted with” includes the implicit provisothat such substitution is in accordance with permitted valence of thesubstituted atom and the substituent, and that the substitution resultsin a stable compound, e.g., which does not spontaneously undergotransformation such as by rearrangement, cyclization, elimination, etc.As used herein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents include acyclic and cyclic, branched andunbranched, carbocyclic and heterocyclic, aromatic and non-aromaticsubstituents of organic compounds. The permissible substituents can beone or more and the same or different for appropriate organic compounds.For purposes of the invention, the heteroatoms such as nitrogen may havehydrogen substituents and/or any permissible substituents of organiccompounds described herein which satisfy the valences of theheteroatoms. Substituents can include any substituents described herein,for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, analkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as athioester, a thioacetate, or a thioformate), an alkoxyl, an alkylthio,an acyloxy, a phosphoryl, a phosphate, a phosphonate, an amino, anamido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl,an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, asulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromaticmoiety.

Unless specifically stated as “unsubstituted,” references to chemicalmoieties herein are understood to include substituted variants. Forexample, reference to an “aryl” group or moiety implicitly includes bothsubstituted and unsubstituted variants.

The term “unsaturated ring” includes partially unsaturated and aromaticrings.

As used herein, the term “tumoral disease” refers to ahyperproliferative disease, such as cancer.

As used herein, the term “conjoint administration” means administrationof two or more agents to a subject of interest as part of a singletherapeutic regimen. The administration(s) can be either simultaneous orsequential, i.e., administering one agent followed by administering of asecond (and/or a third one, etc.) at a later time, as long as the agentsadministered co-exist in the subject being treated, or at least oneagent will have the opportunity to act upon the same target tissues ofother agents while said target tissues are still under the influence ofsaid other agents. In a certain embodiment, agents to be administeredcan be included in a single pharmaceutical composition and administeredtogether. In a certain embodiment, the agents are administeredsimultaneously, including through separate routes. In a certainembodiment, one or more agents are administered continuously, whileother agents are administered only at predetermined intervals (such as asingle large dosage, or twice a week at smaller dosages, etc.).

The present invention includes within its scope the salts and isomers.Compounds of the present invention may in some cases form salts, whichare also within the scope of this invention. The term “salt(s)”, asemployed herein, denotes acidic and/or basic salts formed with inorganicand/or organic acids and bases. Zwitterions (internal or inner salts)are included within the term “salt(s)” as used herein (and may beformed, for example, where the R substituents comprise an acid moietysuch as a carboxyl group). Also included herein are quaternary ammoniumsalts such as alkylammonium salts. Pharmaceutically acceptable (i.e.,non-toxic, physiologically acceptable) salts are preferred, althoughother salts are useful, for example, in isolation or purification stepswhich may be employed during preparation. Salts of the compounds may beformed, for example, by reacting a compound with an amount of acid orbase, such as an equivalent amount, in a medium such as one in which thesalt precipitates or in an aqueous medium followed by lyophilization.

Exemplary acid addition salts include acetates (such as those formedwith acetic acid or trihaloacetic acid, for example, trifluoroaceticacid), adipates, alginates, ascorbates, aspartates, benzoates,benzenesulfonates, bisulfates, borates, butyrates, citrates,camphorates, camphorsulfonates, cyclopentanepropionates, digluconates,dodecylsulfates, ethanesulfonates, fumarates, glucoheptanoates,glycerophosphates, hemisulfates, heptanoates, hexanoates,hydrochlorides, hydrobromides, hydroiodides, 2-hydroxy ethanesulfonates,lactates, maleates, methanesulfonates, 2-naphthalenesulfonates,nicotinates, nitrates, oxalates, pectinates, persulfates,3-phenylpropionates, phosphates, picrates, pivalates, propionates,salicylates, succinates, sulfates (such as those formed with sulfuricacid), sulfonates (such as those mentioned herein), tartrates,thiocyanates, toluenesulfonates, undecanoates, and the like.

Exemplary basic salts (formed, for example, wherein the substituentcomprise an acidic moiety such as a carboxyl group) include ammoniumsalts, alkali metal salts such as sodium, lithium, and potassium salts,alkaline earth metal salts such as calcium and magnesium salts, saltswith organic bases (for example, organic amines) such as benzathines,dicyclohexylamines, hydrabamines, N-methyl-D-glucamines,N-methyl-D-glucamides, t-butyl amines, and salts with amino acids suchas arginine, lysine and the like. The basic nitrogen-containing groupsmay be quaternized with agents such as lower alkyl halides (e.g.,methyl, ethyl, propyl, and butyl chlorides, bromides and iodides),dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl, and diamylsulfates), long chain halides (e.g., decyl, lauryl, myristyl and stearylchlorides, bromides and iodides), aralkyl halides (e.g., benzyl andphenethyl bromides), and others.

Solvates of the compounds of the invention are also contemplated herein.Solvates of the compounds of formula I are preferably hydrates or otherpharmaceutically acceptable solvates.

All stereoisomers of the present compounds, such as those which mayexist due to asymmetric carbons on the R substituents of the compound,including enantiomeric and diastereomeric forms, are contemplated withinthe scope of this invention. Individual stereoisomers of the compoundsof the invention may, for example, be substantially free of otherisomers, or may be admixed, for example, as racemates or with all other,or other selected, stereoisomers. The chiral centers of the presentinvention may have the S or R configuration.

As used herein, the term “treating” or “treatment” includes reversing,reducing, or arresting the symptoms, clinical signs, and underlyingpathology of a condition in manner to improve or stabilize a subject'scondition. As used herein, and as well understood in the art,“treatment” is an approach for obtaining beneficial or desired results,including clinical results. Beneficial or desired clinical results caninclude, but are not limited to, alleviation or amelioration of one ormore symptoms or conditions, diminishment of extent of disease,stabilized (i.e., not worsening) state of disease, preventing spread ofdisease, delay or slowing of disease progression, amelioration orpalliation of the disease state, and remission (whether partial ortotal), whether detectable or undetectable. “Treatment” can also meanprolonging survival as compared to expected survival if not receivingtreatment.

As used herein, a therapeutic that “prevents” a disorder or conditionrefers to a compound that, in a statistical sample, reduces theoccurrence of the disorder or condition in the treated sample relativeto an untreated control sample, or delays the onset or reduces theseverity of one or more symptoms of the disorder or condition relativeto the untreated control sample.

The present application also envisages within its scope the effect ofselection of suitable counterions. The counterion of the compounds ofthe present invention may be chosen by selecting the dissociationconstant for the drug capable of ionization within the said pH range. Byestimating the ionized and un-ionized drug concentration of any compound(using well established equations such a Henderson-Hasselbach equation),the solubility and consequently the absorption of the drug may bealtered.

The compounds generated may be present as a single stereoisomer (e.g.,enriched to at least 95% purity relative to the total amount of allstereoisomers present), a racemate, or a mixture of enantiomers ordiastereomers in any ratio.

Pharmaceutical Compositions

The present invention further provides pharmaceutical compositionscomprising a compound of formula (I) or its pharmaceutically acceptablesalt thereof as an active ingredient along with pharmaceuticallyacceptable additives/excipients/adjuvants/vehicles.

Compounds of the present invention may be used in a pharmaceuticalcomposition, e.g., combined with a pharmaceutically acceptable carrier,for administration to a patient. Such a composition may also containdiluents, fillers, salts, buffers, stabilizers, solubilizers, and othermaterials well known in the art. The term “pharmaceutically acceptable”means a non-toxic material that does not interfere with theeffectiveness of the biological activity of the active ingredient(s).The characteristics of the carrier will depend on the route ofadministration. Such additional factors and/or agents may be included inthe pharmaceutical composition to produce a synergistic effect withcompounds of the invention, or to minimize side effects caused by thecompound of the invention.

The pharmaceutical compositions of the invention may be in the form of aliposome or micelles in which compounds of the present invention arecombined, in addition to other pharmaceutically acceptable carriers,with amphipathic agents such as lipids which exist in aggregated form asmicelles, insoluble monolayers, liquid crystals, or lamellar layers inaqueous solution. Suitable lipids for liposomal formulation include,without limitation, monoglycerides, diglycerides, sulfatides,lysolecithin, phospholipids, saponin, bile acids, and the like.Preparation of such liposomal formulations is within the level of skillin the art, as disclosed, for example, in U.S. Pat. Nos. 4,235,871;4,501,728; 4,837,028; and 4,737,323, all of which are incorporatedherein by reference.

The composition may be administered in a variety of ways includingorally, nasally, buccally, sublingually, intravenously, transmucosally,parenterally, by inhalation, spray, transdermally, subcutaneously,intrathecally, topically or rectally and may be formulated according tomethods known in the art.

The effective dosage form for a mammal may be about 0.1-100 mg/kg ofbody weight of active compound, which may be administered as a singledose or in the form of individual doses, such as from 1 to 4 times aday.

The mammal may be an adult human.

The compounds of the present invention may optionally be administeredwith one or more additional agents. Exemplary additional agents includeone or more compounds independently selected from central nervous systemdrugs, such as CNS/respiratory stimulants, analgesics, narcoticagonists, narcotic antagonists, nonsteroidal anti-inflammatory/analgesicagents, behavior-modifying agents, tranquilizers/sedatives, anestheticagents, inhalants, narcotics, reversal agents, anticonvulsants, skeletalmuscle relaxants, smooth muscle relaxants, cardiovascular agents,inotropic agents, antiarrhythmic drugs, anticholinergics, vasodilatingagents, agents used in treatment of shock, alpha-adrenergic blockingagents, beta-adrenergic blocking agents, respiratory drugs,bronchodilators, sympathomimetics, antihistamines, antitussives, agentsfor urinary incontinence/retention, urinary alkalinizers, urinaryacidifiers, cholinergic stimulants, agents for urolithiasis,gastrointestinal (GI) agents, antiemetic agents, antacids, histamine H2antagonists, gastromucosal protectants, proton pump inhibitors, appetitestimulants, GI antispasmodics-anticholinergics, GI stimulants,laxatives, saline, bulk producing, lubricant, surfactant,antidiarrheals, hormones/endocrine/reproductive agents, sex hormones,anabolic steroids, posterior pituitary hormones, adrenal corticalsteroids, glucocorticoids, antidiabetic agents, thyroid drugs, thyroidhormones, misc. endocrine/reproductive drugs, prostaglandins,antiinfective drugs, antiparasitics, anticoccidial agents, antibiotics,anti-tuberculosis, aminocyclitols, cephalosporins, macrolides,penicillins, tetracyclines, lincosamides, quinolones, sulfonamides,antibacterials, antifungal agents, antiviral agents, blood modifyingagents, clotting agents, anticoagulants, erythropoietic agents,antineoplastics/immunosuppressives, alkylating agents, antidotes,bone/joint agents, dermatologic agents (systemic), vitamins andminerals/nutrients, systemic acidifiers, systemic alkalinizers,anti-cancer agents, and anti-viral agents.

A. Methods of Use

The present invention further provides a method of prophylaxis and/ortreatment of, and/or ameliorating the symptoms of, diseases, comprisingadministering a therapeutically effective amount of a compound offormula (I) or pharmaceutically acceptable salts thereof orpharmaceutical compositions comprising the compound of formula (I) asthe active ingredient.

The compounds of the present invention are useful as c-ABL1 and/orc-ABL2 inhibitors and are useful in all disorders where alteration ofthe amount of c-ABL1 and/or c-ABL2 is required in mammals, includinghumans. The compounds of the present invention may also act as PGDFRaand PGDFRb inhibitors in mammals, including humans. PDGFRa/b isassociated with both cancer (e.g. GIST) as well as cardiovascularabnormalities such as pulmonary arterial hypertension (PAH). Thecompounds of the present invention may also act as inhibitors of stemcell factor receptor (SCFR), also known as c-Kit and mutations in c-Kit,in mammals, including humans. The compounds of the present inventionalso inhibit LCK, and thus may be useful in treating chronic lymphocyticleukemia (CLL).

The compounds of the present invention may be used to treat mammals,including humans, suffering from a tumoral disease a compound of formula(I), e.g., in a therapeutically effective amount.

Imatinib mesylate (Gleevec) has been shown to be effective againstpoxvirus infections by disabling host proteins essential to the viruslife cycle (Nature Medicine, 2005, vol. 11, 7, page 731-739) and withoutinterfering with the acquisition of immune memory (Journal of Virology,2011, vol. 85, 1, p. 21-31).

Similarly, by targeting the host gene products rather the virus itself,administration of imatinib mesylate or nilotinib may be useful intreating Ebola and Marburg virus infections (Science TranslationalMedicine, 2012, vol. 4, 123, page 1-10; Antiviral Research, 2014, vol.106, pages 86-94). Furthermore, Abl family kinases have been shown toregulate the susceptibility of cells to polyomavirus infection bymodulating gangliosides required for viral attachment (Journal ofVirology, 2010, vol. 84, 9, p. 4243-4251). Hence, Abl kinase inhibitor,e.g., a compound of formula (I), may prove useful to treat or prevent apolyomavirus infection.

The present application provides a method for preventing or treating abacterial infection or a viral infection in a subject using a compoundof formula (I) as described herein. In certain embodiments, thebacterial infection is caused by Pseudomonas aeruginosa, Chlamydiatrachomatis, Escherichia coli, Helicobacter pylori, Listeriamonocytogenes, Salmonella typhimurium, Shigella flexneri, orMycobacterium tuberculosis.

In certain embodiments, Mycobacterium tuberculosis causesMDR-tuberculosis or XDR-tuberculosis.

In certain embodiments, the viral infection is caused by a Vacciniavirus, a variola virus, a polyoma virus, a Pox virus, a Herpes virus, acytomegalovirus (CMV), a human immunodeficiency virus, JC virus, JCpolyomavirus (JCV), BK virus, Simian virus 40 (SV40), Monkeypox virus,Ebola virus, Marburg virus, Bunyavirus, Arenavirus, Alphavirus (e.g.,Venezuelan equine encephalitis (VEE), Western equine encephalitis(WEE)), Flavivirus, West Nile virus or Coronavirus (e.g., SARS).

In some embodiments, the compounds described in the present applicationhave improved/maintain desirable safety and toxicity profile relative toimatinib mesylate.

In some embodiments, the compounds described in the present applicationare more soluble than imatinib mesylate in saline and/or at biologicallyuseful pH ranges.

EXEMPLIFICATION Example 1: Synthetic Protocols

Synthesis of(E)-1-(5-bromopyridin-3-yl)-3-(dimethylamino)prop-2-en-1-one (2)

A solution of 1 (40.0 g, 200 mmol) and R-1 (119.0 g, 1000 mmol) in 500mL of THF was stirred at 70° C. overnight. TLC indicated the reactionwas completed. The mixture was cooled to room temperature and removedthe solvent at reduced pressure. The resulting solid was washed withhexane to afford 2 as a yellow solid (47.2 g, 93%).

Synthesis of4-(5-bromopyridin-3-yl)-N-(2-methyl-5-nitrophenyl)pyrimidin-2-amine (3)

A mixture of 2 (45 g, 176.5 mmol), 7 (40.6 g, 159.2 mmol), K₂CO₃ (44.0g, 318.8 mmol) in 500 mL of n-BuOH was heated at 120° C. for 16 hours.The reaction mixture was filtered, and the solvent was removed atreduced pressure. The residue was purified by chromatography column(silica gel, eluted with petroleum ether (PE)/ethyl acetate (EA),PE/EA=2:1) to afford 3 (47.0 g, 70%) was a light yellow solid.

Synthesis ofN¹-(4-(5-bromopyridin-3-yl)pyrimidin-2-yl)-6-methylbenzene-1,3-diamine(4)

A solution of 3 (45.0 g, 116.9 mmol) and SnCl₂ (132.0 g, 585 mmol) in300 mL of EtOAc was heated to reflux overnight, then the reaction wascooled to room temperature, filtered and the solution was concentratedat reduced pressure to afford 4 (44.0 g, 100%). It was directly used forthe next step without any further purification.

Synthesis ofN-(3-(4-(5-bromopyridin-3-yl)pyrimidin-2-ylamino)-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide(5)

The above crude 4 (30.0 g, 84.5 mmol) and 8 (40.0 g, 123.0 mmol) weredissolved in 300 mL of i-BuOH, then the resulting solution was warmed to80° C. for about 5 hours, after completion of the reaction, the mixturewas cooled to room temperature, and removed the solvent under reducedpressure. The resulting residue was purified by flash chromatography onsilica gel (Hexane/EA=2:1) to afford 5 (45.0 g, 93%) as a yellow solid.

Synthesis of5-(2-(2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzamido)phenylamino)pyrimidin-4-yl)pyridin-3-ylboronicacid (6)

A mixture of 5 (10.0 g, 17.5 mmol), KOAc (2.8 g, 28.1 mmol), PCy₃ (0.3g, 1.1 mmol), Pd₂(dba)₃ (0.4 g, 0.5 mmol) and4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (7.1 g, 28.0mmol) in dioxane (150 mL), was stirred at 80° C. overnight, aftercompletion of the reaction. The reaction solution was removed at reducedpressure to afford crude 6 (11.0 g, yield 100%) as yellow solid. It wasdirectly used for next step without further purification.

Synthesis ofN-(3-(4-(5-acetylpyridin-3-yl)pyrimidin-2-ylamino)-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide(7)

A solution of 5 (1.1 g, 2.0 mmol), tributyl(1-ethoxyvinyl)stannane (0.9g, 2.6 mmol), Pd(PPh₃)₄ (0.2 g, 0.1 mmol) and triethylamine (0.3 g, 3.0mmol) in degassed dioxane (50 mL) was heated to reflux for 24 hours. Thesolvent was then evaporated in vacuo and the residue was filteredthrough a thick pad of SiO₂. The solid obtained was taken up in dry THF(60 ml), cooled to 0° C., and treated with 1N HCl. The solution wasstirred for 2 hours at room temperature and then neutralized with sat.aq. NaHCO₃. The mixture was extracted with EA and the organic layerswere washed with brine, dried over Na₂SO₄, and concentrated in vacuo.The residue was purified by flash chromatography (silica gel, elutedwith PE/EA=2:1) to afford 7 (1.0 g, 94%) as a white solid.

Synthesis of ethyl4-(5-(2-(2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzamido)phenylamino)pyrimidin-4-yl)pyridin-3-yl)-2,4-dioxobutanoate(8)

Diethyl oxalate (0.4 g, 2.3 mmol) was added to a suspension of sodiumhydride (70 percent, 0.2 g) in 15 mL of tetrahydrofuran, and refluxedfor about 15 min. Then a solution of 7 (1.0 g, 1.9 mmol) in 5 mL oftetrahydrofuran was added dropwise over 30 min, and refluxed for 90 min.After cooling down, the reaction mixture was poured into ice-cold water,which was neutralized with diluted hydrochloric acid and extracted withethyl acetate, dried over Na₂SO₄, and concentrated. The crude product 8(0.8 g, 67%) was used for the next step without any furtherpurification.

Synthesis of5-(5-(2-(2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzamido)phenylamino)pyrimidin-4-yl)pyridin-3-yl)-4H-pyrazole-3-carboxylicacid (9)

To a solution of 8 (0.8 g, 1.26 mmol) in 20 mL of EtOH, hydrazine (0.1g, 2.54 mmol) was added. The resulting mixture was heated to reflux for60 min. The solution was removed under reduced pressure, and the residuewas purified by prep-HPLC (basic) to afford 9 (0.6 g, 78%) as a whitesolid.

Synthesis of Library Compounds

General Procedure:

A solution of 9 (160 mg, 0.26 mmol), HATU (148 mg, 0.39 mmol), R₁R₂NH(1.2 e.q.) and DIPEA (70 mg, 0.54 mmol) in 2 mL of DMF was stirred for 3hours at room temperature. The resulting mixture was evaporated underreduced pressure and the residue was purified by prep-HPLC to afforddesired library compounds.

Compounds 115, 116 and 117 were prepared using this general procedure.

General Procedure:

A mixture of 5 (200 mg, 0.35 mmol), RB(OH)₂ (2.0 eq.), Pd(PPh₃)₄ (40 mg,0.03 mmol), Na₂CO₃ (112 mg, 1.05 mmol) in dioxane (4 mL) and water (1mL), The resulting reaction mixture was irradiated for 90 min in amicrowave oven. Then the reaction mixture was cooled to room temperatureand concentrated at reduced pressure. The residue was purified byprep-HPLC to afford desired library compounds as a solid.

Compounds 101, 102, 103, 118, 201, 202, 309, 401, 402, 403, 404 and 405were prepared using this general procedure.

General Procedure:

A mixture of 6 (150 mg, 0.34 mmol), RBr or RI (2.0 eq.), Pd(dppf)Cl₂ (57mg, 0.07 mmol), Cs₂CO₃ (280 mg, 0.85 mmol) in i-PrOH (4 mL) and water (1mL) was irradiated for 30 min in a microwave oven. Then the reactionsolution was cooled to room temperature and concentrated at reducedpressure. The residue was purified by prep-HPLC to give desired librarycompounds as a solid.

Compounds 104, 105, 106, 108, 113, 119, 203 were prepared using thisgeneral procedure.

General Procedure:

A solution of 5 (200 mg, 0.35 mmol), R-305, R-306, or R308 (3.0 eq.),Pd₂(dba)₃ (25 mg, 0.03 mmol), t-BuOK (157 mg, 1.40 mmol), BINAP (22 mg,0.03 mmol) in 5 mL of NMP was irradiated for 90 min at 150° C. in amicrowave oven. Then the reaction solution was cooled to roomtemperature and concentrated at reduced pressure. The residue waspurified by prep-HPLC to desired compounds as a solid.

Compounds 305, 306, 308 were prepared using this general procedure.

General Procedures:

To a solution of 5 (200 mg, 0.35 mmol), K₃PO₄ (149 mg, 0.70 mmol), DMCDA(7 mg, 0.05 mmol) and CuI (10 mg, 0.05 mmol) in 2 mL of DMF was addedR-303 or R-304 (2.0 e.q). The resulting mixture was stirred at 120° C.overnight. The reaction mixture was cooled to room temperature, andwater (0.5 mL) was added and extracted with EA (3 mL*3). The combinedorganic layers were dried over Na₂SO₄ and concentrated in vacuo. And theresidue was purified by prep-HPLC to afford desired library compounds asa solid.

Compounds 303 and 304 were prepared using this general procedure.

Synthesis of Compound 107

A solution of 7 (150 mg, 0.28 mmol) in DMF-DMA (3 mL) was stirred at100° C. for 2 hours. Then the solution was cooled to room temperatureand the solvent removed at reduced pressure. The crude residue wasdissolved in EtOH (10 mL), and hydrazine (45 mg, 1.40 mmol) was added.The resulting mixture was heated to reflux overnight. The solvent wascooled to room temperature and concentrated in vacuo. The residue waspurified by prep-HPLC to afford 107 (20 mg, 13%) as a solid.

Synthesis of Compound 207

A solution of 7 (200 mg, 0.37 mmol) in DMA-DMA (4 mL) was heated to 100°C. and stirred for 2 hours. The excess DMA-DMA was evaporated in vacuo,and the residue was dissolved in ethanol (10 mL), to this solution wasadded K₂CO₃ (255 mg, 1.85 mmol) and hydroxylamine hydrochloride (77 mg,1.11 mmol). The resulting mixture was refluxed overnight. After cooling,the mixture was filtered through celite and the filtrate wasconcentrated in vacuo. The residue was purified by prep-HPLC to affordcompound 207 (18 mg, 8%) as a solid.

Synthesis of Compounds 870, 880, 8300, 831, 832

Methods

1-(2-Methyl-5-nitrophenyl)guanidine (1)

A mixture of 2-methyl-5-nitroaniline (152 g, 1.0 mol), cyanamide (247mL, 6.0 mol) and isopropyl alcohol (1000 mL) were placed in a 3 L flask.The mixture was heated to 80° C. Concentrated hydrochloric acid (57 mL)was slowly added dropwise over 80 min. The reaction mixture was stirredfor 1 h while maintaining the temperature at 80° C. Another portion ofconcentrated hydrochloric acid (144 mL) was added dropwise at 80° C. Thereaction mixture was then stirred for 12 h at 100° C. The mixture wascooled to room temperature and treated with aqueous NaOH (2.5N, 1200mL). The resulting solid was collected by filtration, washed withisopropyl alcohol (500 mL) and dried to afford compound 1 (145 g, 76%yield).

(E)-1-(5-Bromopyridin-3-yl)-3-(dimethylamino)prop-2-en-1-one (3)

A mixture of 3-acetyl-5-bromopyridine (126.7 g, 0.633 mol) and DMF-DMA(84 g, 70.6 mmol) was heated under reflux for 1 h. The mixture wascooled to room temperature and then directly purified by silica gelcolumn chromatography. The resulting crude product after concentrationwas washed with diethyl ether (200 mL) and dried to afford compound 3(122 g, 75.5% yield) as yellow crystals.

4-(5-Bromopyridin-3-yl)-N-(2-methyl-5-nitrophenyl)pyrimidin-2-amine (4)

A mixture of compound 1 (10.0 g, 51.5 mmol) and compound 3 (12.9 g, 50.8mmol) in 2-propanol (150 mL) was heated under reflux for 18 h. Themixture was cooled to room temperature and the resulting precipitate wascollected by filtration, washed with diethyl ether (100 mL) and dried toafford compound 4 (13.2 g, 67% yield) as pale yellow crystals.

N¹-(4-(5-Bromopyridin-3-yl)pyrimidin-2-yl)-6-methylbenzene-1,3-diamine(5)

A mixture of iron (5.0 8 g, 907 mmol), NH₄Cl (970 mg, 18.1 mmol) andSiO₂ (3 g) in ethanol/water (1:1, 140 mL) was heated at 55° C. for 10min. Then a suspension of compound 4 (7.0 g, 18.1 mmol) in THF (70 mL)was added. The reaction mixture was stirred under reflux for 1 h andcooled to room temperature. The mixture was poured into water (100 mL)and then extracted with ethyl acetate (100 mL×2). The combined organiclayers were washed with brine and water, dried over anhydrous sodiumsulfate and concentrated to afford compound 5 (5.88 g, 91% yield) asyellow solid.

tert-Butyl 4-(4-(methoxycarbonyl)benzyl)piperazine-1-carboxylate (6a)

TFA (10 mL) was added dropwise to a mixture of methyl 4-formylbenzoate(20 g, 121 mmol) and tert-butyl piperazine-1-carboxylate (25 g, 134mmol) in acetonitrile (400 mL) at room temperature. The mixture wasstirred for 1 h and NaBH₃CN (8.32 g, 134 mmol) was added. The reactionmixture was stirred overnight at room temperature and water was added.The resulting mixture was extracted with ethyl acetate (100 mL×2). Thecombined organic layers were washed with brine and water, dried overanhydrous sodium sulfate and concentrated to afford compound 6a (14 g,34.4% yield), which was used directly in the next step without furtherpurification.

Methyl 4-(piperazin-1-ylmethyl)benzoate (6b)

Compound 6b was prepared from 1-methylpiperazine following the sameprocedure for 6a.

4-((4-(tert-Butoxycarbonyl)piperazin-1-yl)methyl)benzoic acid (7a)

A mixture of compound 6a (7.0 g, crude, 21 mmol) and LiOH—H₂O (1.4 g, 31mmol) in methanol/acetonitrile/water (100 mL, 1:2:2) was stirred 1 h atroom temperature. The organic solvent was removed and the remainingaqueous solution was washed with ethyl acetate (100 mL) and thenadjusted to pH=2-3 with 2N aqueous HCl. The resulting mixture wasextracted with ethyl acetate (30 mL×2). The combined extracts were driedover anhydrous sodium sulfate and concentrated to afford compound 7a(3.0 g, 44.8% yield) as a white solid.

4-(Piperazin-1-ylmethyl)benzoic acid (7b)

Compound 7b was prepared from 6b following the same procedure for 7a

tert-Butyl 4-(4-(3-(4-(5-Bromopyridin-3-yl)pyrimidin-2-ylamino)-4-methylphenylcarbamoyl)benzyl)piperazine-1-carboxylate (8a)

A mixture of compound 5 (1.0 g, 2.81 mmol), compound 7a (0.98 g, 4.19mmol), HATU (1.28 g, 3.37 mmol) in DMF (20 mL) was cooled to 0° C. andDIPEA (1.95 mL, 11.24 mmol) was added. The reaction mixture was allowedto warm to room temperature and stirred for 15 min. Saturated aqueoussodium bicarbonate (20 mL) was added and the resulting mixture wasextracted with ethyl acetate (50 mL×2). The combined extracts were driedover anhydrous sodium sulfate and concentrated. The residue was purifiedby flash column chromatography on silica gel (petroleum ether/ethylacetate=3:1 to 1:1) to afford compound 8a (1.29 g, 80% yield) as ayellow solid.

N-(3-(4-(5-Bromopyridin-3-yl)pyrimidin-2-ylamino)-4-methylphenyl)-4-(piperazin-1-ylmethyl)benzamide(8b)

Compound 8b was prepared from 7b following the same procedure for 8a.

General Procedure for the Final Compounds

8300, 831, 832:

A mixture of compound 8a/b (1.0 eq), the corresponding boronic acid (1.0eq), Pd(dppf)Cl₂ (cat.) and Na₂CO₃ (2.5 eq) in 1-4-dioxane and water(5:1) was stirred at 80° C. for 1 h under N₂. The mixture was cooled toroom temperature and then diluted with ethyl acetate and water. Theresulting mixture was filtered and the filtrate was separated. Theaqueous phase was extracted with ethyl acetate. The combined organicphases were washed with brine, dried over anhydrous sodium sulfate andconcentrated to dryness. The residue was purified by flash columnchromatography on silica gel or prep-HPLC to afford compound of interestas a yellow solid.

870:

To a mixture of compound 8a (100 mg, 0.152 mmol), pyridin-4-ylboronicacid (21 mg, 0.167 mmol), Pd(dppf)Cl₂ (15 mg, cat.) and Na₂CO₃ (40 mg,0.608 mmol) in 1-4-dioxane (2.5 mL) and water (0.5 mL) was stirred at80° C. for 1 h under N₂. The mixture was cooled to room temperature andthen diluted with ethyl acetate (10 mL) and water (10 mL). The resultingmixture was filtered and the filtrate was separated. The aqueous phasewas extracted with EtOAc (20 mL×2). The combined organic phases werewashed with brine (20 mL×2), dried over anhydrous and concentrated todryness. The residue was purified by flash column chromatography onsilica gel (DCM/MeOH=100:1 to 20:1) to afford compound Boc-870 (100 mg)as a brown solid.

TFA (1 mL) was added to a solution of Boc-870 (100 mg, 0.152 mmol) inCH₂Cl₂ (4 mL) at 0° C. with stirring. The reaction mixture was stirredfor 1 h and concentrated to dryness. The residue was treated withaqueous NaHCO₃ to adjust pH=9 and then extracted with ethyl acetate (5mL×3). The combined organic layers were washed with water and brine,dried over anhydrous sodium sulfate and concentrated. The residue waspurified by flash column chromatography on silica gel (DCM/MeOH=20:1 to10:1) to afford compound 870 (66 mg, 78.4% yield) as an off-white solid.

880:

Compound 880 was prepared from pyridin-2-ylboronic acid following thesame procedure for 870.

Synthesis of 810, 820, 830, 840, 8150, 8170, 8190, 8200, 8220, 8250,8260, 8270, 8280, 8290

Methods

General Procedure for Compound 9 from Compound 4

810, 820, 830, 840

To a mixture of compound 4 (1.0 eq), the corresponding boronic acid (1.0eq), Pd(dppf)Cl₂ (cat.) and Na₂CO₃ (2.5 eq) in 1-4-dioxane and water(5:1) was stirred at 80° C. for 1 h under N₂. The mixture was cooled toroom temperature and then diluted with ethyl acetate and water. Theresulting mixture was filtered and the filtrate was separated. Theaqueous phase was extracted with ethyl acetate. The combined organicphases were washed with brine, dried over anhydrous sodium sulfate andconcentrated to afford compound 9, which was used in the next stepwithout further purification.

N-(2-Methyl-5-nitrophenyl)-4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)pyrimidin-2-amine(11)

A mixture of compound 4 (5.0 g, 12.95 mmol), bis(pinacolato)diboron(3.62 g, 14.25 mmol), Pd(dppf)Cl₂ (0.3 g, cat.) and KOAc (3.83 g, 38.87mmol) in toluene (50 mL) was heated under reflex for 12 h under N₂. Themixture was cooled to room temperature and diluted with ethyl acetate(100 mL) and water (100 mL). The resulting mixture was filtered and thefiltrate was separated. The aqueous phase was extracted with ethylacetate (100 mL×2). The combined organic phases were washed with brine(30 mL), dried over anhydrous sodium sulfate and concentrated todryness. The residue was purified by flash column chromatography onsilica gel (Petroleum/EtOAc=20:1) to afford compound 11 (4.77 g, 84.3%yield) as a brown solid.

General Procedure for Compound 9 from Compound 11

8150, 8170, 8190, 8200, 8220, 8250, 8260, 8270, 8280, 8290

A mixture of Ar-X (1.0 eq), compound 11 (1.1 eq), Pd(dppf)Cl₂ (cat.) andNa₂CO₃ (3.0 eq) in 1-4-dioxane and water (5:1) was stirred at 80° C. for1 h under N₂. The mixture was cooled to room temperature and dilutedwith ethyl acetate and water. The resulting mixture was filtered and thefiltrate was separated. The aqueous phase was extracted with ethylacetate and the combined organic phases were washed with brine, driedover anhydrous sodium sulfate and concentrated to afford compound 9,which was used in the next step without further purification.

General Procedure for Compound 10

A mixture of iron (5.0 eq), NH₄Cl (1.0 eq) and SiO₂ (2.0 eq) inethanol/water (1:1) was heated at 55° C. for 10 min. Then a suspensionof compound 9 (1.0 eq) in THF was added. The reaction mixture wasstirred under reflux for 1 h and cooled to room temperature. The mixturewas poured into water and then extracted with ethyl acetate. Thecombined organic layers were washed with brine and water, dried overanhydrous sodium sulfate and concentrated to afford compound 10 as ayellow solid.

General Procedure for the Final Compound

A mixture of compound 10 (1.0 eq), compound 7b (1.2 eq) and HATU (1.2eq) in DMF (20 mL) was cooled to 0° C. and DIPEA (4.0 eq) was added. Thereaction mixture was allowed to warm to room temperature and stirred for15 min. Saturated aqueous sodium bicarbonate was added and the resultingmixture was extracted with ethyl acetate. The combined extracts weredried over anhydrous sodium sulfate and concentrated. The residue waspurified by flash column chromatography on silica gel to afford thedesired compound compound as a solid.

Synthesis of 806, 809, 8120, 8130, 8180, 8230, 8140

4-Methoxy-N-(2-methyl-5-nitrophenyl)pyrimidin-2-amine (12)

To a mixture of 2-chloro-4-methoxypyrimidine (9.54 g, 66 mmol),2-methyl-5-nitrobenzenamine (10.0 g, 66 mmol), Pd₂(dba)₃ (1.0 g), S-Phos(1.0 g, 24.4 mmol), and Cs₂CO₃ (31.8 g, 99 mmol) in 1,4-dioxane/water(140 mL/60 mL) was heated at 110° C. overnight. The mixture was cooledto room temperature and then filtered through a pad of celite. Thefiltrate was diluted with ethyl acetate and washed with water. Theorganic phase was dried over anhydrous sodium sulfate and concentrated.The residue was purified by flash column chromatography on silica gel toafford compound 12 (12 g. 70.6% yield) as a light yellow solid.

2-(2-Methyl-5-nitrophenylamino)pyrimidin-4-ol (13)

A mixture of compound 12 (20 g, 77 mol), TMSCl (15 g, 136 mmol) and NaI(23.4 g, 156 mmol) in acetonitrile (400 mL) was heated at 120° C.overnight. The mixture was cooled to room temperature and 2N aqueousNa₂CO₃ (400 mL) and DCM (400 mL) were added. The organic layer wasseparated, washed with water, dried over anhydrous sodium sulfate andconcentrated. The residue was purified by flash column chromatography onsilica gel (DCM/MeOH=200:1) to afford compound 13 (12.1 g, 64% yield) asa light yellow solid.

4-Chloro-N-(2-methyl-5-nitrophenyl)pyrimidin-2-amine (14)

A mixture of compound 13 (2 g, 8.13 mmol) and DMF (5 drops) in POCl₃ (40mL) was heated under reflux for 2 h. The mixture was cooled to roomtemperature and most of POCl₃ was removed. The residue was poured intoaqueous NaOH (100 mL) carefully and the resulting mixture was extractedwith DCM (100 mL×2). The combined organic layers were washed with brineand water (100 mL), dried over anhydrous sodium sulfate and concentratedto afford compound 14 (2.0 g, 93% yield) as a yellow solid.

General Procedure for Compound 15

A mixture of compound 14 (1.0 eq), PTC-1480-X-Int (1.0 eq), Pd(dppf)Cl₂(cat.) and K₂CO₃ (3.0 eq) in 1-4-dioxane (20 mL) and water (20 mL) washeated under reflux for 12 h under N₂. The mixture was cooled to roomtemperature and diluted with ethyl acetate and water. The resultingmixture was filtered and the filtrate was separated. The aqueous phasewas extracted with ethyl acetate. The combined organic phases werewashed with brine, dried over anhydrous sodium sulfate and concentratedto dryness. The residue was purified by flash column chromatography onsilica gel (DCM/MeOH=200:1) to afford compound 15 as a yellow solid.

General Procedure for Compound 16

To a mixture of iron (1.0 eq), NH₄Cl (2.0 eq) and SiO₂ (cat) inethanol/water (1:1) was heated at 55° C. for 10 min. Then a suspensionof compound 15 (2.0 eq) in THF was added. The reaction mixture wasstirred under reflux for 1 h and cooled to room temperature. The mixturewas poured into water and then extracted with ethyl acetate. Thecombined organic layers were washed with brine and water, dried overanhydrous sodium sulfate and concentrated to afford compound 16.

Procedure for 806

A mixture of compound 16 (Het=3-methylisoxazol-5-yl, 150 mg, 0.42 mmol),compound 7a (134 mg, 0.42 mmol) and HATU (159 mg, 0.42 mmol) in DMF (2mL) was cooled to 0° C. and DIPEA (217 mg, 1.68 mmol) was added. Thereaction mixture was allowed to warm to room temperature and stirred for15 min. Saturated aqueous sodium bicarbonate was added and the resultingmixture was extracted with ethyl acetate. The combined extracts weredried over anhydrous sodium sulfate and concentrated. The residue waspurified by flash column chromatography on silica gel (Petroleumether/ethyl acetate=1:1 to ethyl acetate) to afford compound Boc-806(180 mg, 65.2% yield) as a solid.

HCl/EtOAc (2N, 1 mL) was added to a solution of Boc-806 (97 mg, 0.147mmol) in EtOAc (1 mL) at 0° C. with stirring. The reaction mixture wasstirred for 1 h and the resulting precipitate was collected byfiltration, washed with DCM and dried to afford compound 806 (HCl salt,80 mg, 100% yield) as a yellow solid.

General Procedure 806, 809, 8120, 8130, 8180, 8230 and 8240

A mixture of compound 16 (1.0 eq), compound 7b (1.0 eq) and HATU (1.0eq) in DMF (2 mL) was cooled to 0° C. and DIPEA (4.0 eq) was added. Thereaction mixture was allowed to warm to room temperature and stirred for15 min. Saturated aqueous sodium bicarbonate was added and the resultingmixture was extracted with ethyl acetate. The combined extracts weredried over anhydrous sodium sulfate and concentrated. The residue waspurified by flash column chromatography on silica gel (DCM/MeOH=20:1) toafford final compound as a yellow solid.

Example 2: Inhibition of Abelson Protein Kinases c-Abl1, c-Abl2 andc-Kit and Comparison to Imatinib, the Active Ingredient in Gleevec®

Cpd Abl1 (nM) Abl2 (nM) c-Kit (nM) 101 232.35 224.57 6.80 102 95.33169.62 6.70 103 200.41 212.18 7.70 107 45.17 66.95 7.20 108 40.53 85.887.40 113 35.00 50.11 7.50 114 101.53 142.60 32.40 115 107.81 202.79 8.90116 91.06 160.89 11.60 117 29.13 34.99 5.70 118 37.16 51.00 8.40 119117.66 44.68 4.00 201 193.18 514.16 19.30 202 403.88 701.00 31.10 203250.56 711.96 47.10 207 30.19 73.73 12.20 303 162.15 349.36 18.90 305397.83 631.35 9.20 300 174.37 149.18 14.00 401 167.28 178.63 12.40 402183.14 207.42 11.10 404 100.32 116.17 5.8o 405 107.09 150.43 5.90 806 84152 13 809 47 77 7.8 820 84 173 10 830 34 51 6.0 832 53 77 4.0 880 203341 12 8120 476 783 27 8130 323 423 44 8170 128 182 11 8180 369 303 138190 97 91 9.7 8200 96 131 4.3 8230 >1000 >1000 37 8240 >1000 >1000 398250 71 216 15 8260 41 236 16 8270 53 100 4.9 8280 57 104 6.2 8290 51137 5.7 8300 40 39 5.2 imatinib 828.3 1000 30.7Measurement of c-Abl1, c-Abl2 and c-Kit IC50 Values

Kinase base buffer (50 mM HEPES, pH 7.5 0.0015% Brij-35; 10 mM MgCl₂ 2mM DTT) and Stop buffer

(100 mM HEPES, pH 7.5 0.015% Brij-35; 0.2% Coating Reagent (50 mM EDTA)are prepared. Test compound is diluted in 100% DMSO to 50-times thedesired final inhibitor concentration (the Stock Solution) and seriallydiluted in half-log increments resulting in final concentrations 250 μMto 75 μM, 25 μM, 7.5 μM, 2.5 μM, 0.75 μM, 0.25 μM, 75 nM, 25 nM, 7.5 nMin DMSO. 10 μl of each compound is placed in a 96-well plate as theintermediate plate. 90 μl of Kinase Buffer is added to to each well toprepare the intermediate plate.

Mix the compounds in intermediate plate for 10 min on shaker. For theassay of enzyme inhibitions, 5 μl of each well from the intermediateplate is transferred to a 384-well plate in duplicates, 10. Then 10 μlof 2.5× enzyme solution is added to each well of the 384-well assayplate and incubated for 10 min. Then enzyme substrate is added as 10 μlof 2.5× FAM-labeled peptide+ATP solution to each well of the 384-wellassay plate The reaction is allowed to proceed at 28° C. and quenchedwith the addition of 25 μl of stop buffer. The release of fluorescentFAM is quantitated as Percent inhibition=(max-conversion)/(max-min)*100.

“max” stands for DMSO control; “min” stands for low control. Data arefit in XLFit excel add-in version 4.3.1 to obtain IC50 values. Equationused is: Y=Bottom+(Top−Bottom)/(1+(IC50/X)^HillSlope)

Example 3: Inhibition Profile of a 500 nM Solution of Test CompoundsAgainst 14 Protein Kinases

Kinase base buffer (50 mM HEPES, pH 7.5 0.0015% Brij-35; 10 mM MgCl₂ 2mM DTT) and Stop buffer

(100 mM HEPES, pH 7.5 0.015% Brij-35; 0.2% Coating Reagent (50 mM EDTA)are prepared. Test compound is diluted in 100% DMSO to 50-times thedesired final inhibitor concentration (the Stock Solution) in DMSO. 10μl of each compound is placed in a 96-well plate as the intermediateplate. 90 μl of Kinase Buffer is added to to each well to prepare theintermediate plate. Mix the compounds in intermediate plate for 10 minon shaker. For the assay of enzyme inhibitions, 5 μl of each well fromthe intermediate plate is transferred to a 384-well plate in duplicates,10. Then 10 μl of 2.5× enzyme solution is added to each well of the384-well assay plate and incubated for 10 min. Then enzyme substrate isadded as 10 μl of 2.5× FAM-labeled peptide+ATP solution to each well ofthe 384-well assay plate The reaction is allowed to proceed at 28° C.and quenched with the addition of 25 μl of stop buffer. The release offluorescent FAM is quantitated as Percentinhibition=(max−conversion)/(max−min)*100.

“max” stands for DMSO control; “min” stands for low control. Convertconversion values to inhibition values.Percent inhibition=(max−conversion)/(max−min)*100.“max” stands for DMSO control; “min” stands for low control.

TABLE 1 PDG PDG ARG/ Cpd YES FRa LCK SRC ABL FLT3 KIT FRb FGR LYNA Abl2FES FYN JNK2 101 100 42 88 18 73 36 87 99 55 89 74 43 52 25 102 99 39 9119 83 32 99 99 67 88 75 42 55 26 103 101 42 91 25 71 28 92 98 60 91 7138 54 17 107 101 59 95 28 91 51 99 101 82 93 87 28 69 1.7 108 100 56 9434 91 73 100 101 80 93 88 67 66 36 113 100 61 94 37 92 25 98 101 85 9591 26 75 18 114 100 34 77 18 84 20 99 99 81 91 78 17 72 18 115 100 53 9029 83 22 96 99 78 93 76 20 67 17 116 100 48 94 29 87 8.4 93 101 83 93 7619 67 5.9 117 99 74 97 55 94 13 95 99 90 97 91 24 86 14 118 99 66 95 3592 61 93 101 84 96 88 38 77 14 119 99 67 98 53 83 90 95 100 86 97 90 4782 42 201 99 45 91 18 72 30 91 98 67 87 55 49 53 6.1 202 100 46 87 16 6419 85 98 64 87 50 27 47 19 203 100 29 88 17 72 31 77 95 59 82 54 21 4317 207 100 54 95 30 94 19 88 99 79 92 85 31 72 19 303 100 33 87 13 79 2587 100 58 86 62 31 45 4.7 305 100 16 80 10 62 20 98 100 47 80 50 47 254.4 309 101 25 84 13 77 13 96 99 59 87 79 31 44 24 401 100 40 88 18 7624 98 99 67 90 81 19 50 23 402 100 29 86 19 72 31 96 100 65 89 75 23 4821 404 100 47 91 21 82 24 97 100 75 90 83 32 56 34 405 101 33 90 16 8029 95 100 68 90 80 33 52 28

TABLE 2 Cpd Abl2 Abl1 PDGFR^(a) PDGFR^(b) JNK1 JNK2 SRC LCK CKIT FES YESFYN LYNA FLT3 FGR 810 59 65 99 98 25 48 20 83 85 23 44 71 93 26 57 82076 85 99 98 12 29 23 88 87 32 56 73 91 61 64 830 87 92 100 99 23 52 3893 92 22 63 81 97 44 77 831 60 48 100 97 15 17 12 82 82 23 25 57 87 3834 832 81 88 100 100 24 16 28 93 91 24 66 79 106 58 80 840 30 38 99 9224 56 15 50 73 35 25 64 77 69 24 870 60 60 100 97 17 37 17 70 82 39 2358 83 73 38 880 61 71 100 100 28 48 15 70 87 35 33 64 80 62 46 8150 5442 99 97 31 42 14 77 83 26 26 62 90 68 34 8170 73 78 100 98 32 40 25 8886 67 49 70 93 79 68 8190 81 82 100 99 41 58 28 92 86 51 52 74 96 72 718200 75 78 98 97 41 54 22 92 85 52 50 65 100 68 63 8220 54 44 99 97 5650 22 90 83 57 40 75 93 75 47 8250 68 87 100 98 15 8 20 87 80 37 45 7485 68 63 8260 67 85 100 101 21 15 22 83 79 16 41 71 88 62 60 8270 80 9199 98 41 36 33 91 88 60 60 83 104 78 81 8280 80 90 100 99 23 14 28 82 8932 53 76 98 54 78 8290 75 86 100 100 33 23 26 87 88 36 55 76 98 63 758300 89 92 99 98 35 30 45 97 88 19 63 90 108 61 81

Example 4: JCV Antiviral Potency and Therapeutic Index for SelectedCompounds

TABLE 3 Fold Improvement Anti-JCV potency in Therapeutic Com- relativeto imatinib Index pound (EC₅₀ ^(Imatinib):EC₅₀ ^(Novel))(TI^(Novel):TI^(Imatinib)) 102 1.9 0.71 103 7.7 >3.3 107 2.4 0.4 1080.71 0.15 113 0.52 0.14 114 Inactive — 117 Inactive — 118 <0.1 <0.15 20732.7 >13.3 309 2.7 1 809 40 >167 830 1.5 >6.3 832 2.2 >9.2 8190 9.4 >398250 0.79 >3.3 8270 5.7 >24 8280 11.2 >47 8290 2.7 >11.3 imatinib 1 1

EC₅₀ was measured in triplicate with a multiplicity of infection of 0.02over 7 days in Cos7 cells using the laboratory strain MAD-1, originallyderived from the brain of PML patients. EC₅₀ was computed using PRISMand quality of fit regression coefficients >0.85 for each case. Relativeantiviral potency was computed relative to the EC₅₀ of imatinib.Relative therapeutic index was computed by measuring the cellcytotoxicity CC₅₀:EC₅₀ ratio to calculate the Therapeutic Index (TI)with drug titration between 2 nM and 20 μM.

With respect to compound 118, only an upper limit to EC₅₀ could becalculated because 118 had little antiviral activity and poor qualitydata fit. With respect to compounds 103, 207, 809, 830, 832, 8190, 8250,8270 and 8280, only the lower limit of relative TI could be calculatedbecause the drug substance showed no toxicity up to 20 μM. The absolutevalue of EC₅₀ for imatinib was measured as 4.91 μM.

Example 5: Anti-Cancer Potency of Selected Compounds Against Mutantc-Kit Proteins Associated with Gastrointestinal Stromal Tumor

96 well plates were created using an HP D300 programmable digitaldispensing tool (Hewlett-Packard). 10 mM drug stocks in DMSO weretitrated into each well and each schematic plate map was recorded. Dosesranged from 3 nM to 1000 nM, depending upon the drug and experiment.Imatinib was used as a control drug and loaded on the plate in the sameconcentrations as for the test compounds. Media with DMSO at the highestconcentration of the compounds was used as a negative control. Thesepre-made plates with dispensed drug and media were then frozen at −20°C. until used. Plates were warmed before cells were loaded. GIST celllines with different genotypes were plated at 2000 cells per well in a100 μl total volume. The plated cells were incubated for 72 hours, 5%CO₂, and 37° C. Proliferation/viability was measured usingCellTiter-Glo® luminescent cell viability reagent which is based on thequantitation of ATP present in a cell. Plate was then read using GloMax®96 microplate luminometer. Results are depicted in FIGS. 1A-1G.

Example 6: Western Blot Demonstrating that Selected CompoundsAnti-Cancer Response is Due to a Block in Formation of Phosphorylatedc-Kit (p-Kit)

Confluent T-25 flasks of GIST-related c-Kit protein containing the557-558 deletion were expressed in T1 cells were treated with testcompound for 90 minutes, 5% CO₂, 37° C. Untreated and imatinib treatedcells served as controls. After lysis, 175 μg of whole cell lysate wassubject to protein electrophoresis and transferred to nitrocellulose.KIT phosphorylation was visualized by probing the blot with P-KIT (Y719)antibody. AKT phosphorylation was visualized by probing with P-AKT. MAPKphosphorylation was visualized by probing with P-MAPK. After blot wasstriped, it was re-probed for total protein with KIT CD117 antibody, AKTantibody, and MAPK antibody, respectively. The blot is shown in FIG. 2.

Example 7: Anti-Cancer Potency of Selected Compounds Against MutantBCR-Abl Proteins Associated with Chronic Myelogenous Leukemia

TABLE 4 IC50 (Ba/F3; nM) Pa- Native rental p210 T315I Y253H E255K E255VF359V 103 2923 20.23 2906 2594 196.2 1196 135.3 113 2065 12.17 2124 143446.89 738.6 161 207 2801 15.69 2796 2379 116.2 1290 128.8 820 2484 60.042303 2774 203.7 2670 260.3 860 4637 281.4 3429 7061 857 3130 792.8 8906035 31.72 6553 5328 64.24 1447 213.4 8190 2237 58.54 2677 2098 389.7997.2 236.5 8270 2521 18.82 2789 2269 45.9 1030 203.3 8280 2614 66.962770 2458 234.4 2389 251.4 8290 2583 98.75 2483 2872 80.4 1444 375.18300 2994 29.46 3775 1444 251.1 614.4 148.3 832 2485 9.939 2590 158490.86 661.1 46.7 Imatinib 213Methods

The “parental” cell line refers to untransduced Ba/F3 cells grown in thepresence of IL-3. The IC50s of the compounds against these cells arelisted. To measure the IC50 values, pools of Ba/F3 cells harboring thespecified isoform of BCR-ABL were established following retroviraltransduction with MSCV puro BCR-ABL constructs and selected in thepresence of puromycin. IL-3 was subsequently withdrawn from thesepopulations.

For the experiment, exponentially growing cells were plated at a finalconcentration of 50,000 cells/mL in 10% RPMI containing 0.5% DMSO in atotal volume of 100 μL per well of a 96-well opaque plate. Finalconcentrations of each compound were 10 μM, 1 μM, 100 nM, 10 nM, 1 nM,0.1 nM, 0 nM (plus a blank—media only). Each cell line and concentrationwas plated in triplicate for this single experiment. Plates wereincubated for 48 hrs at 37° C. and read with Cell Titer Glo on a platereader. Data was compiled with the Sensimax software; raw values werenormalized to the untreated; averages were taken of the three normalizedreplicate wells and IC50 values were calculated via Prism 5.

Example 8: Suppression of Tumor Growth for K562 Cell BCR-Abl Xenograftin NSG Mice as a Model of Chronic Myelogenous Leukemia

Methods:

5×10⁶ K562 cells derived from a patient with CML and stably expressingthe BCR-Abl transgene product were implanted subcutaneously in thehindlimb of NSG female mice, 7-9 weeks of age. The tumor was allow todevelop to approximately 100 mm³ prior to initiation of once per day(Q.D.) dosing at 0 (i.e. vehicle), 15 mg/kg, 25 mg/kg or 40 mg/kg forcompound 832 by oral gavage in 50 mM sodium citrate as a vehicle, pH3.5. Dosing was initiated at approximately day 6 after implantation andcontinued for 9 days. Measurement of tumor volume, veterinaryobservations and animal weight changes were used to determine theeffectiveness of the drug with 5 mice per group. The results aredepicted in FIG. 3.

All publications and patents cited herein are hereby incorporated byreference in their entirety.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

We claim:
 1. A compound having the following structure, or apharmaceutically acceptable salt thereof:


2. The compound of claim 1, wherein the compound is a methanesulfonicacid salt.
 3. The compound of claim 1, wherein the compound is asuccinic acid salt.
 4. The compound of claim 1, wherein the compound isa free base.
 5. A pharmaceutical composition comprising the compound ofclaim 1 and a pharmaceutically acceptable excipient.
 6. A pharmaceuticalcomposition comprising the compound of claim 2 and a pharmaceuticallyacceptable excipient.
 7. A pharmaceutical composition comprising thecompound of claim 3 and a pharmaceutically acceptable excipient.
 8. Apharmaceutical composition comprising the compound of claim 4 and apharmaceutically acceptable excipient.